This document discusses various diagnostic techniques for viral animal diseases. It describes direct detection methods like electron microscopy, histopathology, and fluorescent antibody techniques. It also covers indirect detection methods like ELISA, immunochromatography, latex agglutination, and viral antibody detection techniques like complement fixation, haemagglutination inhibition, and virus neutralization tests. Emerging techniques discussed include PCR, microarrays, and nanobiosensor-based diagnostics.
The lecture was presented to the students of Saudi board of Community Medicine to help them know about the various serological methods applicable in the diagnosis of infectious diseases in general with attention upon the specificity and sensitivity of various diagnostic modalities. The lecture covers the basic principles of each test and the clinical applications with the advantages and disadvantages of each.
SEROLOGICAL DIAGNOSTIC TEST AND IT'S INTERPRETATION FOR INFECTIOUS DISEASESfahadzubairi
A presentation SEROLOGICAL DIAGNOSTIC TEST AND IT'S INTERPRETATION FOR INFECTIOUS DISEASES.
Dr. Sayed Inseram Ali. B.Sc. ( Hons ), M.Sc.; AACC, ASMT, ASM, : USA; R.M. CANADA; IFCC U. K.
The lecture was presented to the students of Saudi board of Community Medicine to help them know about the various serological methods applicable in the diagnosis of infectious diseases in general with attention upon the specificity and sensitivity of various diagnostic modalities. The lecture covers the basic principles of each test and the clinical applications with the advantages and disadvantages of each.
SEROLOGICAL DIAGNOSTIC TEST AND IT'S INTERPRETATION FOR INFECTIOUS DISEASESfahadzubairi
A presentation SEROLOGICAL DIAGNOSTIC TEST AND IT'S INTERPRETATION FOR INFECTIOUS DISEASES.
Dr. Sayed Inseram Ali. B.Sc. ( Hons ), M.Sc.; AACC, ASMT, ASM, : USA; R.M. CANADA; IFCC U. K.
Immunological diagnosis of parasitic infectionRaghwendra sah
This slide is just made for the students who want note and don't get access to the book which cost more. So, hope you all will get information about the immunological diagnosis of parasitic infection.
Lab report that discusses the antigen-antibody precipitation reaction using the Ouchterlony Double Diffusion Technique.
Created by: Annisa Hayatunnufus
Bachelor of Pharmacy
Management & Science University
Immunological diagnosis of parasitic infectionRaghwendra sah
This slide is just made for the students who want note and don't get access to the book which cost more. So, hope you all will get information about the immunological diagnosis of parasitic infection.
Lab report that discusses the antigen-antibody precipitation reaction using the Ouchterlony Double Diffusion Technique.
Created by: Annisa Hayatunnufus
Bachelor of Pharmacy
Management & Science University
serological techniques for detection of plant virus.pptxReddykumarAv
Serological tests involve diagnostic procedures for identifying antibodies and antigens in a patient's blood sample. Serology definition tells that Serological tests could be used to diagnose infections and autoimmune disorders, as well as to see whether a person is resistant to these kinds of diseases and for a variety of other purposes, including assessing a person's blood type.
LABORATORY DIAGNOSIS OF VIRAL INFECTIONS.pdfWani Insha
Laboratory diagnosis of viral infections is useful for the following purposes:
To start antiviral drugs for those viral infections for which specific drugs are available such as herpes, CMV, HIV, influenza and respiratory syncytial virus (RSV)
Screening of blood donors for HIV, hepatitis B and hepatitis C-helps in prevention of transfusion transmitted infections
Surveillance purpose: To assess the disease burden in the community by estimating the prevalence and incidence of viral infections
For outbreak or epidemic investigation, e.g. influenza epidemics, dengue outbreaks-to initiate appropriate control measures
To start post-exposure prophylaxis of antiretroviral drugs to the health care workers following needle stick injury.
To initiate certain measures: For example,
If rubella is diagnosed in the first trimester of pregnancy, termination of pregnancy is recommended
If newborn is diagnosed to have hepatitis B infection, then immunoglobulins (HBIG) should be started within 12 hours of birth.
This presentation explores a brief idea about the structural and functional attributes of nucleotides, the structure and function of genetic materials along with the impact of UV rays and pH upon them.
Multi-source connectivity as the driver of solar wind variability in the heli...Sérgio Sacani
The ambient solar wind that flls the heliosphere originates from multiple
sources in the solar corona and is highly structured. It is often described
as high-speed, relatively homogeneous, plasma streams from coronal
holes and slow-speed, highly variable, streams whose source regions are
under debate. A key goal of ESA/NASA’s Solar Orbiter mission is to identify
solar wind sources and understand what drives the complexity seen in the
heliosphere. By combining magnetic feld modelling and spectroscopic
techniques with high-resolution observations and measurements, we show
that the solar wind variability detected in situ by Solar Orbiter in March
2022 is driven by spatio-temporal changes in the magnetic connectivity to
multiple sources in the solar atmosphere. The magnetic feld footpoints
connected to the spacecraft moved from the boundaries of a coronal hole
to one active region (12961) and then across to another region (12957). This
is refected in the in situ measurements, which show the transition from fast
to highly Alfvénic then to slow solar wind that is disrupted by the arrival of
a coronal mass ejection. Our results describe solar wind variability at 0.5 au
but are applicable to near-Earth observatories.
Comparing Evolved Extractive Text Summary Scores of Bidirectional Encoder Rep...University of Maribor
Slides from:
11th International Conference on Electrical, Electronics and Computer Engineering (IcETRAN), Niš, 3-6 June 2024
Track: Artificial Intelligence
https://www.etran.rs/2024/en/home-english/
What is greenhouse gasses and how many gasses are there to affect the Earth.moosaasad1975
What are greenhouse gasses how they affect the earth and its environment what is the future of the environment and earth how the weather and the climate effects.
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
2. substances or devices which aid in the diagnosis of viral infections
Classification of viral diagnostic tests
(i) direct virus detection
(ii) viral antigen detection
(iii) virus isolation
(iv) viral antibody detection.
Viral diagnostics
3. Electron microscopy, immunoelectron microscopy
Histopathology
Fluorescent antibody technique – direct, indirect
Immunohistochemistry
ELISA
Immunochromatography
Latex agglutination test
Direct identification of viruses
4. ELECTRON MICROSCOPY
Rapid diagnosis
No need of additional probes
Disadvantage
Procurement and maintenance of the instrument
– expensive
Require skilled & well trained personnel for
operation.
Sensitivity is comparatively low – require at least
10 6 viral particles for + ve diagnosis.
Single specimen examined at a time.
Applications:
Rotavirus from faecal samples, rabies virus from
brain, norwalk virus from faecal samples
5. Employs viral antibody
Increases the sensitivity & specificity of EM
Two methods (i) classical IEM (ii) solidphase IEM
classical IEM: Requires prior incubation of the clinical sample with virus
specific antibody .
Solidphase IEM: solid support (copper grid) is coated with specific antibody
which captures viruses from the clinical sample.
Immunoelectron microscopy (IEM)
6. Helps in diagnosis of viral inclusion bodies
Viral inclusion bodies: Aggregates of viral nucleocapsids in the cytoplasm or
nucleus.
Can be detected by H&E staining
Applications
Negri bodies in rabies virus
Guarnieri bodies – vaccinia virus
Bollinger bodies - fowlpox
Histopathology
7. Use of two specific antibodies – nanoparticle conjugated antibody, antibody
fixed in chromatography paper.
The sample dropped on sample pad – forms complex with the antibody. When
moves along the membrane pad- sandwiched between two antibodies-
production of colour.
diagnosis of avian influenza, dengue etc.
IMMUNOCHROMATOGRAPHY
8. Particulate antigen +virus specific antibody - crosslinking of polyvalent
antigens – agglutination.
Visibility can be improved by coating the latex beads with virus specific
antibodies (sensitized latex beads).
Sensitized latex beads + clinical sample (containing viruses) visible
aggregates.
Applications:
Avian influenza
Rota virus
FMD (Sugimura et al.,2000)
Latex agglutination test
9. Fluorescence antibody technique (FAT)
Fluorochromes like fluorescent isothiocyanate and rhodamine isothiocyanate. Are
used.
Viral antigens can be detected from smears from clinical samples, frozen tissue
sections and also from formalin fixed tissue samples.
Direct Fluorescence antibody technique
The virus specific antibodies are directly tagged with fluorescent dyes.
Indirect fluorescence antibody technique (IFAT)
second antibody known as anti-species immunoglobulin (Igs) are used.
Advantage:
Available easily commercially than the virus specific antibodies
More sensitivity than direct FAT.
Better signal amplification
10. Immunohistochemistry
Principle
Employs antibodies conjugated with enzymes like horseradish peroxidase or
alkaline phosphatase.
When suitable substrates are used, colour reaction takes place in case of
positive sample which can be viewed through light microscope.
Both direct and indirect immunoperoxidase tests are used.
Advantages
The procedure can be applied for formalin fixed tissues as well.
Comparative study of site of localization of viruses and the tissue damage is
possible
11. Sandwich ELISA
The viral antigen is sandwiched between two antibodies namely capture and
detection antibodies. For the assay both the antibodies should target different
epitopes of the antigen. This is followed by addition of enzyme conjugated anti-
species immunoglobulin. On adding suitable chromogen-substrate colour
reaction develops.
Advantages
Assay is quantitative, amount of viral antigen can be detected
Assay has high sensitivity and specificity
More samples can be tested at the same time
Disadvantages
Need ELISA reader for result interpretation; not possible under field conditions
The method is time consuming and labourious.
Applications:
Used in diagnosis of PPR, Bluetongue, FMD etc
13. Complement fixation test
Principle
The ability of the complement system to fix the antigen- antibody complex
forms the basis of complement fixation test.
Sheep RBCs and its corresponding antibody acts as indicator system.
When the antigen reacts with specific antisera, antigen antibody complex will
be formed and the complement will be unavailable for the indicator system.
Then there will be no haemolysis and the test is positive.
If the antiserum is not specific, then the complement is free to fix the
indicator system resulting in haemolysis.
Sensitive assay like ELISA have replaced it.
Simple; easy to perform.
Applications
Complement fixation test is prescribed by the OIE for diagnosis of equine
diseases like African horse sickness, equine encephalomyelitis (eastern,
western, Venezuelan), vesicular stomatitis.
14. Haemagglutination inhibition tests
Principle
Certain viruses namely paramyxoviruses and orthomyxoviruse have the ability to
bind to sialic acid residues in RBCs. This property is called haemagglutination.
If specific antibody and viruses are mixed prior to the addition of RBCs
haemagglutination is inhibited.
HI test is used for serotyping and also for measuring antibody titre.
Applications
OIE recommends this test for diagnosis of viral infections like blue tongue,
avian laryngeo tracheitis, avian influenza, marek’s disease, infectious bursal
disease, enzootic bovine leucosis, equine infectious anaemia (Coggin’s test),
myxomatosis and caprine arthritis encephalitis disease.
16. Virus neutralization test
Principle
When a serum sample to be tested is specific for the virus, there will be
antigen-antibody complex formation. The virus particles are unavailable to
cause cytopathic effect in cell culture which can be visualized microscopically
or by staining with vital dyes.
Applications
Prescribed test by OIE for diagnosis of pseudorabies, rabies, Riftvalley fever,
vesicular disease and IBRT-IPV.
17. Plaque reduction neutralization test
Principle
The ability of virus specific antibody to inhibit the plaque forming property of
viruses by 50% when plated in semisolid media is the basic principle involved.
Applications
PRNT is recommended by the OIE for diagnosis of eastern equine encephalitis
and Venezulean equine encephalitis disease.
Disadvantages:
The interpretation of results requires at least 3-5 days which is very slow when
compared to EIA and RIA.
Both the tests involve use of cell culture where false results due to
contamination of cell culture are possible.
18. Agar gel immunodiffusion test
Principle
It is basically a precipitation test in which soluble antigen reacts with its
homologous antibody in the presence of electrolytes. At the optimum
proportion, an insoluble antigen-antibody precipitate will be formed.
Applications
OIE recommends this test for diagnosis of viral infections like blue tongue,
avian laryngeo tracheitis, avian influenza, marek’s disease, infectious bursal
disease, enzootic bovine leucosis, equine infectious anaemia (Coggin’s test),
myxomatosis and caprine arthritis encephalitis disease.
19. Counter immunoelectrophoresis
PRINCIPLE
Counter immunoelectrophoresis is the immunodiffusion modified by
electrophoresis to drive antigen and antibody towards each other. The specimen
to be tested is placed in the cathode side, and the antiserum is placed in the
anode. At neutral or alkaline pH, antigens are negatively charged and hence
migrate towards anode in barbital or veronal buffer.
Applications:
Counter immunoelectrophoresis is used in diagnosis of blue tongue virus, orf
virus and pox viral diseases
20. Radio immunoassay
Principle
It is the competitive binding assay which binding of known amount of
radiolabeled substrate and antibody. When the test sample is added, the
radiolabeled substrate gets displaced by it resulting in free radiolabelled
substrate in the solution. Finally, the radioactivity of free substrate in the
solution is measured which is proportional to the amount of unknown antigen
bound to antibody.
The assay is more sensitive and specific compared to the above methods.
Disadvantage:
Assay needs sophisticated instruments like gamma counter.
Radioactive compounds are hazardous to human health ie. Carcinogenic.
Therefore, it is not routinely used for viral diagnosis.
21. Indirect ELISA
PRINCIPLE
The viral antigen is immobilized in a solid support followed by addition of
serially diluted antibodies. The antibodies in turn are captured by enzyme
conjugated anti-species immunoglobulin.
Addition of chromogen / substrate facilitates the colour reaction which can
be measured calorimetrically.
Currently, use of recombinant viral proteins as antigens reduce the risk of
handling of dangerous organisms and made the assay sensitive and specific.
Advantages
Simplicity; easy to perform
Rapid than neutralization assays
Safer: Use of enzymes instead of radiolabels
Require ELISA reader for result interpretation
23. RFLP
The nucleotide signatures of each species are unique. This fact is explored in
the technique where endonucleases recognizing restriction sites are used to cut
the genome at various sites for each organism. The genome fragments are
resolved by running in agarose gel electrophoresis. The characteristic nucleotide
base pair length corresponds to each organism.
27. Nanobiosensor
There are various types of nanobiosensors based on various
principles namely, electrochemical biosensors, voltametric and
amperometric sensors, impedance sensors, optical fiber based
sensors, surface plasmon resonance based biosensors, quartz
crystal microbalance and atomic force microscopy based
nanobiosensors.
Nanobiosensors have been designed for diagnosis of dreadful
human viral diseases like HIV, hepatitis B, Hepatitis C, Ebola virus
etc. Nanobiosensors have also been designed for diagnosis of
animal diseases like avian influenza virus, infectious bovine
rhinotracheitis, rabies, bovine leukemia virus.